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(Mouse) Shh Antibody (N-term) 精选

Purified Rabbit Polyclonal Antibody (Pab)

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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Hela (Human Cervical epithelial adenocarcinoma cell line) cells labeling Shh with AP21229a at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-rabbit IgG (NK179883) secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm and membrane staining on Hela cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin (PD18466410) at 1/100 dilution (red).The nuclear counter stain is DAPI (blue).
AP21229a staining (Mouse) Shh in mouse cerebellum sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Overlay histogram showing HT-29 cells stained with AP21229a (green line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP12735b, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit lgG (H+L) (1583138) at 1/400 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Anti-Shh Antibody (N-term)at 1:2000 dilution + mouse stomach lysates Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution Predicted band size : 48 kDa Blocking/Dilution buffer: 5% NFDM/TBST.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	FC, IHC-P, IF, WB, E
Primary Accession	Q62226
Reactivity	Human, Rat, Mouse
Host	Rabbit
Clonality	polyclonal
Isotype	Rabbit IgG
Calculated MW	47773 Da

Additional Information
Gene ID	20423
Target/Specificity	This mouse Shh antibody is generated from a rabbit immunized with a KLH conjugated synthetic peptide between 58-91 amino acids from the N-terminal region of mouse Shh.
Dilution	FC~~1:25 IHC-P~~1:100~500 IF~~1:25 WB~~1:2000 E~~Use at an assay dependent concentration.
Format	Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	(Mouse) Shh Antibody (N-term) is for research use only and not for use in diagnostic or therapeutic procedures.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	Shh {ECO:0000303\|PubMed:7916661, ECO:0000312\|MGI:MGI:98297}
Function	[Sonic hedgehog protein]: Precursor of sonic hedgehog, a morphogen that activates the smoothened signaling pathway, and which is essential for a variety of patterning events during development (PubMed:11389830, PubMed:31657721, PubMed:7736596, PubMed:7891723, PubMed:8824192). The C-terminal part of the precursor displays an autoproteolysis and a cholesterol transferase activity, resulting (1) in the cleavage of the full-length protein into two parts, Sonic hedgehog protein N-product and C-product (ShhN and ShhC, respectively) and (2) covalent attachment of a cholesterol moiety to the C-terminus of the newly generated ShhN (PubMed:7736596, PubMed:7891723, PubMed:8824192). Both autoproteolysis and a cholesterol transferase activities occur in the endoplasmic reticulum (PubMed:21357747). Following additional lipidation, ShhN acts as a morphogen, while ShhC is degraded in the endoplasmic reticulum (PubMed:21357747).
Cellular Location	[Sonic hedgehog protein]: Endoplasmic reticulum membrane {ECO:0000250\|UniProtKB:Q15465}. Golgi apparatus membrane {ECO:0000250\|UniProtKB:Q15465}. Note=Co-localizes with HHAT in the ER and Golgi membrane. {ECO:0000250\|UniProtKB:Q15465}
Tissue Location	Expressed in a number of embryonic tissues including the notochord, ventral neural tube, floor plate, lung bud, zone of polarizing activity and posterior distal mesenchyme of limbs (PubMed:7916661). In the adult, expressed in lung and neural retina (PubMed:7916661).